CN113773374A - 转录因子ZmbZIPa6及其编码基因与应用 - Google Patents
转录因子ZmbZIPa6及其编码基因与应用 Download PDFInfo
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- CN113773374A CN113773374A CN202010499200.1A CN202010499200A CN113773374A CN 113773374 A CN113773374 A CN 113773374A CN 202010499200 A CN202010499200 A CN 202010499200A CN 113773374 A CN113773374 A CN 113773374A
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Abstract
本发明公开了转录因子ZmbZIPa6及其编码基因与应用。本发明首先公开了一种蛋白质,所述蛋白质为氨基酸序列如SEQ ID NO.2所示的蛋白质或SEQ ID NO.2所示的氨基酸序列的N端或/和C端连接蛋白标签得到的融合蛋白。本发明进一步提供了上述蛋白质的编码基因及其在调控植物耐旱性中的应用。本发明将ZmbZIPa6基因导入野生型拟南芥中,得到转ZmbZIPa6拟南芥与野生型拟南芥相比,其耐旱性及耐碱性明显提高,说明ZmbZIPa6是与植物耐旱和耐碱相关的基因;对于培育耐旱性及耐碱性的作物、林草等新品种具有重要的理论及实际意义,可用于农牧业和生态环境治理所需的抗性植物品种的培育与鉴定。
Description
技术领域
本发明属于生物技术领域,具体涉及转录因子ZmbZIPa6及其编码基因与应用。
背景技术
中国气候复杂,气象灾害种类多、频率高,干旱、盐碱、极端温度等非生物胁迫是引起全球粮食作物减产的首要原因,造成了许多重要经济作物大量减产,而干旱是主要气象灾害之一。这些非生物胁迫对植物可能造成伤害,面对各种非生物胁迫,植物虽不能像动物一样逃离胁迫环境,但也会对胁迫信号做出相应响应,通过一系列生理生化上、细胞和分子水平上的反应来应对不利环境。
充分发掘鉴定植物中与胁迫相关的响应基因,利用分子生物学等技术来研究植物对干旱、盐碱和低温等一系列逆境的应答途径及基因表达调控分子机制,以及利用基因工程提高作物的抗逆性,在理论研究及实际生产上具有重要意义。
发明内容
本发明所要解决的技术问题为如何有效提高植物的耐逆性,尤其是耐旱性和耐碱性。
为解决上述技术问题,本发明首先提供了一种蛋白质,名称为ZmbZIPa6蛋白或蛋白质ZmbZIPa6,来源于玉米(Zea mays),为如下A1)或A2)或A3)任一所示:
A1)氨基酸序列如SEQ ID NO.2所示的蛋白质;
A2)SEQ ID NO.2所示的氨基酸序列的N端或/和C端连接蛋白标签得到的融合蛋白;
A3)将SEQ ID NO.2所示的氨基酸序列经过一个或几个氨基酸残基的取代和/或缺失和/或添加得到的与A1)所示的蛋白质具有90%以上的同一性且功能相同的蛋白质。
其中,SEQ ID NO.2由324个氨基酸残基组成。
上述蛋白质可人工合成,也可先合成其编码基因,再进行生物表达得到。
上述蛋白质中,蛋白标签(protein-tag)是指利用DNA体外重组技术,与目的蛋白一起融合表达的一种多肽或者蛋白,以便于目的蛋白的表达、检测、示踪和/或纯化。所述蛋白标签可为Flag标签、His标签、MBP标签、HA标签、myc标签、GST标签和/或SUMO标签等。
上述蛋白质中,同一性是指氨基酸序列的同一性。可使用国际互联网上的同源性检索站点测定氨基酸序列的同一性,如NCBI主页网站的BLAST网页。例如,可在高级BLAST2.1中,通过使用blastp作为程序,将Expect值设置为10,将所有Filter设置为OFF,使用BLOSUM62作为Matrix,将Gap existence cost,Per residue gap cost和Lambda ratio分别设置为11,1和0.85(缺省值)并进行检索一对氨基酸序列的同一性进行计算,然后即可获得同一性的值(%)。
上述蛋白质中,所述90%以上的同一性可为至少91%、92%、95%、96%、98%、99%或100%的同一性。
上述蛋白质的编码基因也在本发明的保护范围之内。
本发明上述蛋白质的编码基因可为如下B1)或B2)或B3)中任一所示:
B1)SEQ ID NO.1第413-1387位所示的DNA分子;
B2)编码序列为SEQ ID NO.1第413-1387位所示的DNA分子;
B3)在严格条件下与B1)或B2)限定的DNA分子杂交,且编码蛋白质ZmbZIPa6的DNA分子。
其中,SEQ ID NO.1由1648个核苷酸组成,其编码序列为SEQ ID NO.1所示第413-1387位,编码氨基酸序列如SEQ ID NO.2所示的蛋白。
所述严格条件是在2×SSC,0.1%SDS的溶液中,在68℃下杂交并洗膜2次,每次5min,又于0.5×SSC,0.1%SDS的溶液中,在68℃下杂交并洗膜2次,每次15min。
含有上述蛋白质的编码基因的表达盒、重组载体、重组微生物、转基因植物细胞系、转基因植物组织、转基因植物器官或转基因植株也在本发明的保护范围之内。
上文中,所述表达盒是含有编码上述蛋白质的核酸分子的表达盒(即ZmbZIPa6基因表达盒),指能够在宿主细胞中表达蛋白质ZmbZIPa6的DNA分子,该DNA分子不但可包括启动ZmbZIPa6基因转录的启动子,还可包括终止ZmbZIPa6基因转录的终止子。进一步,所述表达盒还可包括增强子序列。可用于本发明的启动子包括但不限于:组成型启动子,组织、器官和发育特异的启动子和诱导型启动子。启动子的例子包括但不限于:花椰菜花叶病毒的组成型启动子35S;来自西红柿的创伤诱导型启动子,亮氨酸氨基肽酶("LAP",Chao等人(1999)Plant Physiol 120:979-992);来自烟草的化学诱导型启动子,发病机理相关1(PR1)(由水杨酸和BTH(苯并噻二唑-7-硫代羟酸S-甲酯)诱导);西红柿蛋白酶抑制剂II启动子(PIN2)或LAP启动子(均可用茉莉酮酸曱酯诱导);热休克启动子(美国专利5,187,267);四环素诱导型启动子(美国专利5,057,422);种子特异性启动子,如谷子种子特异性启动子pF128(CN101063139B(中国专利2007 1 0099169.7)),种子贮存蛋白质特异的启动子(例如,菜豆球蛋白、napin,oleosin和大豆beta conglycin的启动子(Beachy等人(1985)EMBO J.4:3047-3053)。它们可单独使用或与其它的植物启动子结合使用。此处引用的所有参考文献均全文引用。合适的转录终止子包括但不限于:农杆菌胭脂碱合成酶终止子(NOS终止子)、花椰菜花叶病毒CaMV 35S终止子、tml终止子、豌豆rbcS E9终止子和胭脂氨酸和章鱼氨酸合酶终止子(参见,例如:Odell等人(I985)Nature313:810;Rosenberg等人(1987)Gene,56:125;Guerineau等人(1991)Mol.Gen.Genet,262:141;Proudfoot(1991)Cell,64:671;Sanfacon等人Genes Dev.,5:141;Mogen等人(1990)Plant Cell,2:1261;Munroe等人(1990)Gene,91:151;Ballad等人(1989)Nucleic Acids Res.17:7891;Joshi等人(1987)Nucleic Acid Res.,15:9627)。
在本发明具体的实施方式中,所述表达盒为SEQ ID NO.3所示的ZmbZIPa6基因表达盒,包括CaMV35Sq启动子、ZmbZIPa6基因的编码序列和pA35S终止子,其中,CaMV35Sq启动子为SEQ ID NO.3自5’末端第1-698位、ZmbZIPa6基因的编码序列为SEQ ID NO.3自5’末端第1128-2102位(对应SEQ ID NO.1自5’末端第413-1387位)、pA35S为SEQ ID NO.3自5’末端第2107-2325位。
上文中,所述重组载体可含有SEQ ID NO.1第413-1387位所示的用于编码蛋白质ZmbZIPa6的DNA分子。
可用植物表达载体构建含有所述ZmbZIPa6基因表达盒的重组载体。所述植物表达载体可为Gateway***载体或双元农杆菌载体等,如pLeela、pGWB411、pGWB412、pGWB405、pBin438、pCAMBIA1300、pCAMBIA super1300、pCAMBIA1302、pCAMBIA2300、pCAMBIA2301、pCAMBIA1301、pBI121、pCAMBIA1391-Xa或pCAMBIA1391-Xb。使用ZmbZIPa6构建重组载体时,在其转录起始核苷酸前可加上任何一种增强型、组成型、组织特异型或诱导型启动子,如花椰菜花叶病毒(CAMV)35S启动子、泛生素基因Ubiqutin启动子(pUbi)等,它们可单独使用或与其它的植物启动子结合使用;此外,使用本发明的基因构建植物表达载体时,还可使用增强子,包括翻译增强子或转录增强子,这些增强子区域可以是ATG起始密码子或邻接区域起始密码子等,但必需与编码序列的阅读框相同,以保证整个序列的正确翻译。所述翻译控制信号和起始密码子的来源是广泛的,可以是天然的,也可以是合成的。翻译起始区域可以来自转录起始区域或结构基因。
为了便于对转基因植物细胞或植物进行鉴定及筛选,可对所用植物表达载体进行加工,如加入可在植物中表达的编码可产生颜色变化的酶或发光化合物的基因(GUS基因、萤光素酶基因等)、具有抗性的抗生素标记物(庆大霉素标记物、卡那霉素标记物等)或是抗化学试剂标记基因(如抗除莠剂基因)等。
在本发明具体的实施方式中,所述重组载体为pLeela-ZmbZIPa6。所述pLeela-ZmbZIPa6是将植物表达载体pLeela的attR1和attR2重组位点间的序列替换为SEQ ID NO.3所示的DNA分子,且保持其它序列不变得到的载体;其中,SEQ ID NO.3所示的DNA分子为ZmbZIPa6基因表达盒,包括CaMV35Sq启动子、ZmbZIPa6基因编码序列和pA35S终止子,其中,CaMV35Sq启动子为SEQ ID NO.3自5’末端第1-698位、ZmbZIPa6基因的编码序列为SEQ IDNO.3自5’末端第1128-2102位(对应SEQ ID NO.1自5’末端第413-1387位)、pA35S终止子为SEQ ID NO.3自5’末端第2107-2325位。
上文中,所述重组微生物具体可为酵母、细菌、藻和真菌;如所述细菌可以为农杆菌Gv3101。
上文中,所述转基因植物器官可为转基因植物的根、茎、叶、花、果实和种子。
上文中,所述转基因植物细胞系、转基因植物组织和转基因植物器官均不包括繁殖材料。
本发明进一步提供了上述蛋白质、上述蛋白质的编码基因、含有上述蛋白质的编码基因的表达盒、重组载体、重组微生物、转基因植物细胞系、转基因植物组织、转基因植物器官或转基因植株在下述任一种中的应用:
C1)调控植物耐旱性;
C2)制备调控植物耐旱性的产品;
C3)提高植物耐旱性;
C4)制备提高植物耐旱性的产品;
C5)调控植物耐碱性;
C6)制备调控植物耐碱性的产品;
C7)提高植物耐碱性;
C8)制备提高植物耐碱性的产品;
C9)调控植物对脱落酸(ABA)的敏感性;
C10)制备调控植物对ABA的敏感性的产品;
C11)提高植物对ABA的敏感性;
C12)制备提高植物对ABA的敏感性的产品。
上述蛋白质、上述蛋白质的编码基因、含有上述蛋白质的编码基因的表达盒、重组载体、重组微生物、转基因植物细胞系、转基因植物组织、转基因植物器官或转基因植株在植物育种中的应用也在本发明的保护范围之内。
上述应用中,所述植物育种中的应用具体可为将含有上述蛋白质ZmbZIPa6或上述蛋白质ZmbZIPa6的编码基因的植物与其它植物进行杂交以进行植物育种。
本发明进一步提供了提供一种培育耐旱性和/或耐碱性高的转基因植物的方法。
本发明培育耐旱性和/或耐碱性高的转基因植物的方法,包括提高目的植物中上述蛋白质ZmbZIPa6的基因的表达量和/或所述蛋白质ZmbZIPa6的含量和/或所述蛋白质ZmbZIPa6的活性,得到转基因植物;所述转基因植物的耐旱性和/或耐碱性高于所述目的植物。
上述方法中,所述提高目的植物中蛋白质ZmbZIPa6的基因的表达量和/或所述蛋白质ZmbZIPa6的含量和/或所述蛋白质ZmbZIPa6的活性的方法为在目的植物中表达或过表达蛋白质ZmbZIPa6。
上述方法中,所述表达或过表达的方法为将蛋白质ZmbZIPa6的编码基因导入目的植物。
上述方法中,将蛋白质ZmbZIPa6的编码基因导入目的植物可通过携带有本发明ZmbZIPa6基因的植物表达载体导入目的植物中。携带有本发明ZmbZIPa6的植物表达载体可通过使用农杆菌介导、Ti质粒、Ri质粒、植物病毒载体、直接DNA转化、微注射、电导等常规生物学方法转化植物细胞或组织,并将转化的植物细胞或组织培育成植株。
上述方法中,所述蛋白质ZmbZIPa6的编码基因的核苷酸序列是SEQ ID NO.1第413-1387位所示的DNA分子。
在本发明具体的实施方式中,所述携带有本发明ZmbZIPa6的植物表达载体可为pLeela-ZmbZIPa6。所述pLeela-ZmbZIPa6是将植物表达载体pLeela的attR1和attR2重组位点间的序列替换为SEQ ID NO.3所示的DNA分子,且保持其它序列不变得到的载体。
本发明中耐旱性具体体现在:a)通过PEG胁迫作用下转基因植物的存活率大于所述目的植物;b)通过土壤干旱胁迫作用下转基因植物的存活率高于所述目的植物。
本发明中耐碱性具体体现在通过pH9.0胁迫作用下转基因植物的生长情况要明显好于所述目的植物。
本发明中,所述植物是如下E1)-E4)中的任一种:
E1)双子叶植物;
E2)单子叶植物;
E3)十字花科植物;
E4)禾本科植物;
其中,所述十字花科植物如拟南芥(Arabidopsis thaliana);所述禾本科植物如玉米(Zea mays)。
本发明的实验证明从玉米中筛选到一个受ABA诱导表达的ZmbZIPa6基因,将该基因导入野生型拟南芥中,得到转ZmbZIPa6拟南芥与野生型拟南芥相比,该转ZmbZIPa6拟南芥的耐旱性及耐碱性明显提高,说明ZmbZIPa6是与耐旱性和耐碱性相关的蛋白,参与胁迫响应。因此ZmbZIPa6是与植物耐旱和耐碱相关的基因;对于培育耐旱性及耐碱性的作物、林草等新品种具有重要的理论及实际意义,可用于农牧业和生态环境治理所需的抗性植物品种的培育与鉴定。
附图说明
图1为转ZmbZIPa6拟南芥株系的表达量鉴定(A)及正常生长条件下的生长情况(B)。
图2为转ZmbZIPa6拟南芥的ABA胁迫下萌发期的生长情况(A)和根长的比较情况(B)。
图3为转ZmbZIPa6拟南芥的ABA胁迫下苗期的生长情况;其中,A为含10μM的1/2MS培养基苗期生长情况;B为含20μM ABA的1/2MS培养基苗期生长情况;C为含50μM ABA的1/2MS培养基苗期生长情况。
图4为转ZmbZIPa6拟南芥PEG模拟干旱胁迫下苗期生长情况(A)和根长的比较情况(B)。
图5为转ZmbZIPa6拟南芥土壤干旱胁迫下生长情况。
图6为转ZmbZIPa6拟南芥碱胁迫下生长情况(A)和根长的比较情况(B)。
图1-6中,WT代表野生型拟南芥Col-0,1-6代表转ZmbZIPa6拟南芥株系1-6,2-4代表转ZmbZIPa6拟南芥株系2-4,3-3代表转ZmbZIPa6拟南芥株系3-3。
具体实施方式
下面结合具体实施方式对本发明进行进一步的详细描述,给出的实施例仅为了阐明本发明,而不是为了限制本发明的范围。以下提供的实施例可作为本技术领域普通技术人员进行进一步改进的指南,并不以任何方式构成对本发明的限制。
下述实施例中的实验方法,如无特殊说明,均为常规方法。下述实施例中所用的材料、试剂等,如无特殊说明,均可从商业途径得到。
下述实施例中的定量试验,均设置三次重复实验,结果取平均值。
下述实施例中用到的主要材料和试剂的来源如下:
玉米自交系B73,记载于非专利文献“Zea mays L.;ZmbZIP60mRNA is spliced inmaize in response to ER stress,BMC Research Notes(2012)5:144.”,公众可从中国科学院植物研究所获得,以重复本申请实验,不可作为其它用途使用。
RNase抑制剂(购自Takara公司),反转录酶SuperScript TM III(购自Invitrogen公司),pENTR/D-TOPO(购自Invitrogen公司),Phusion High-Fidelity DNA Polymerase(购自NEB公司)。
植物表达载体pLeela记载于非专利文献“A novel role for histonemethyltransferase KYP/SUVH4in the control of Arabidopsis primary seeddormancy,New Phytologist(2012)193:605-616”,公众可从中国科学院植物研究所获得,以重复本申请实验,不可作为其它用途使用。
农杆菌Gv3101(pMP90RK)(Agrobacterium tumefaciens strain GV3101(pMP90RK))记载于非专利文献“Binary Agrobacterium vectors for planttransformation,M Bevanin,Nucleic Acids Research(1984)12:8711-8721.”,公众可从中国科学院植物研究所获得,以重复本申请实验,不可作为其它用途使用。
拟南芥Col-0(ecotype columbia,Arabidopsis thaliana)记载于非专利文献“Arabidopsis,a useful weed.Meyerowitz EM,Cell(1989)56:263-270.”,公众可从中国科学院植物研究所获得,以重复本申请实验,不可作为其它用途使用。
实施例1、ZmbZIPa6基因的获得
提取三叶一心期的玉米自交系B73的叶片的总RNA并以其为模板进行反转录;其中,反转录反应体系为:
(1)DNA消化:10ul体系
条件:37℃30min
(2)反转录:25ul体系
充分解链,加入经步骤(1)DNA消化的总RNA 10ul
OligodT 1ul
DEPC-H20 2.5ul
条件:70℃温育5min,冰上放置5min。
条件:42℃温育1h
以步骤(2)反转录得到的cDNA稀释10倍作为模板,利用引物5'-TGCCACTGAGTGTGTGCATGATT-3'和引物5'-ACTCAAAAGGCAGCCGAACC-3'及Phusion High-Fidelity DNA Polymerase进行PCR扩增,获得PCR产物,其中,反应条件为:先98℃预变性45sec,然后98℃变性10sec,54℃退火20sec,再72℃延伸1.5min,共40个循环;最后72℃延伸10min。
将PCR产物进行琼脂糖凝胶电泳检测,结果得到大小为1008bp左右的PCR产物;回收该PCR产物条带。
将该PCR产物与载体pENTR/D-TOPO连接,获得中间载体pENTR-ZmbZIPa6;所述pENTR-ZmbZIPa6是将pENTR/D-TOPO的第569-804位替换为PCR产物,且保持其他序列不变得到的载体。
将SEQ ID NO.1所示的基因命名为ZmbZIPa6,经过测序,该PCR产物的序列如SEQID NO.1自5’末端第382-1389位所示,包含SEQ ID NO.1自5’末端第413-1387位所示的ZmbZIPa6的编码序列,将该基因编码的蛋白命名为ZmbZIPa6,该蛋白的氨基酸序列为SEQID NO.2。
实施例2、转ZmbZIPa6拟南芥的获得及其功能研究
一、转ZmbZIPa6拟南芥的获得
1、重组载体pLeela-ZmbZIPa6的获得
将实施例1构建的中间载体pENTR-ZmbZIPa6通过LR反应(LR克隆酶,购自Invitrogen公司)将实施例1的PCR产物导入植物表达载体pLeela,同源重组得到重组载体。
经过测序,该重组载体为将植物表达载体pLeela的attR1和attR2重组位点间的序列替换为SEQ ID NO.3所示的DNA分子,且保持其它序列不变得到的载体,该重组载体受35S启动子驱动,将该重组载体命名为pLeela-ZmbZIPa6。
其中,SEQ ID NO.3所示的DNA分子为ZmbZIPa6基因表达盒,包括CaMV35Sq启动子、ZmbZIPa6基因的编码序列和pA35S终止子,其中,CaMV35Sq启动子为SEQ ID NO.3自5’末端第1-698位、ZmbZIPa6基因的编码序列为SEQ ID NO.3自5’末端第1128-2102位(对应SEQ IDNO.1自5’末端第413-1387位)、pA35S终止子为SEQ ID NO.3自5’末端第2107-2325位。
2、重组菌的获得
将步骤1得到的重组载体pLeela-ZmbZIPa6转化到农杆菌Gv3101(pMP90RK)细胞中,用含有50μg/ml硫酸卡那霉素、50μg/ml庆大霉素、50μg/ml利福平、50μg/mL羧苄霉素的YEB培养基进行筛选,得到重组菌。
提取重组菌的质粒送去测序,结果该质粒为pLeela-ZmbZIPa6,说明为阳性重组菌,将该重组菌命名为Gv3101(pMP90RK)/pLeela-ZmbZIPa6。
3、转ZmbZIPa6拟南芥的获得及筛选
1)转ZmbZIPa6拟南芥的获得
将步骤2得到的重组菌Gv3101(pMP90RK)/pLeela-ZmbZIPa6采用花序浸泡法转化野生型拟南芥Col-0,得到T0代转ZmbZIPa6拟南芥。
2)转ZmbZIPa6拟南芥的筛选
取T0代转ZmbZIPa6拟南芥种子播种于含有100μg/ml羧苄青霉素钠的MS培养基上,将具有抗性的成活幼苗移至温室培养(培养温度22℃,光周期16/8小时),收集15株T1代转ZmbZIPa6拟南芥种子并播种于含100μg/ml羧苄青霉素钠的MS培养基中,选取3株分离比为3:1的T1代转ZmbZIPa6拟南芥的成活幼苗(5-10棵)移至温室培养,收集T2代转ZmbZIPa6拟南芥种子并播种于含100μg/ml羧苄青霉素钠的MS培养基中,不发生分离转基因拟南芥即纯合体。最终选取T2代转ZmbZIPa6拟南芥纯合体株系1-6、2-4、3-3进行下述试验。
3)检测转基因拟南芥中ZmbZIPa6的表达
提取3周苗龄的T2代转ZmbZIPa6拟南芥纯合体株系1-6、2-4、3-3和野生型拟南芥Col-0的总RNA,并反转录得到的cDNA稀释10倍作为模板(方法同实施例1),利用引物F:5'-GTCTTTATACAACCTTACCCTCG-3'和引物R:5'-GCACCATCTGAATCTACACCAT-3'行PCR扩增;并以Actin1基因为内参,引物为Actin1-F:5-'CATCAGGAAGGACTTGTACGG-3',Actin1-R:5'-GATGGACCTGACTCGTCATAC-3'。
其中,反应体系(10ul)如下:
94℃预变性5min,94℃变性30sec,55℃退火30sec,72℃延伸20sec,共35个循环;最后再72℃延伸10min。
检测结果如图1中A所示,野生型拟南芥Col-0(WT)中没有扩增出目的条带,转ZmbZIPa6拟南芥株系1-6、2-4、3-3中皆扩增出了目的条带,说明ZmbZIPa6在这3个转ZmbZIPa6拟南芥株系中均表达,而野生型拟南芥Col-0没有ZmbZIPa6表达,说明转ZmbZIPa6拟南芥株系1-6、2-4、3-3为阳性。
观察植株的生长情况,结果如图1中B所示,发现未经任何处理的正常生长条件下的转ZmbZIPa6拟南芥株系1-6、2-4、3-3与野生型拟南芥Col-0(WT)的生长情况无明显区别,可用于进一步的耐逆性等相关研究。
二、转ZmbZIPa6拟南芥耐逆性研究
1、转ZmbZIPa6拟南芥ABA敏感性鉴定
将步骤一中T2代转ZmbZIPa6拟南芥株系1-6、2-4、3-3和野生型拟南芥Col-0(WT)的种子直接播在0.5μM ABA的1/2MS培养基上竖直培养3周,结果如图2中A所示,转ZmbZIPa6拟南芥株系1-6、2-4、3-3的生长显著受到抑制,部分种子仅能露出胚;对根长进行比较,结果如图2中B所示,转ZmbZIPa6拟南芥株系1-6、2-4、3-3的根长均明显短于野生型拟南芥Col-0(WT),表明转ZmbZIPa6拟南芥株系在种子萌发后期ABA呈现高敏感性。
同时将T2代转ZmbZIPa6拟南芥株系1-6、2-4、3-3和野生型拟南芥Col-0(WT)种子播于1/2MS培养基上,4℃春化3天后,在22℃、50%湿度、光照16h和黑暗8h的条件下培养,将生长3d的幼苗分别移到10μM,20μM,50μM ABA的1/2MS培养基上,竖直培养3周,检测了苗期对ABA的敏感性,结果如图3所示,发现含10μM(图3中A),20μM ABA(图3中B)的1/2MS培养基生长的转ZmbZIPa6拟南芥株系1-6、2-4、3-3根明显短于野生型拟南芥Col-0(WT),且叶子发黄;含50μM ABA的1/2MS培养基上的转ZmbZIPa6拟南芥株系1-6、2-4、3-3几乎不能生长,植株死亡率比野生型拟南芥Col-0(WT)高(图3中C)。
上述结果表明,转ZmbZIPa6基因的拟南芥株系对高浓度的ABA较为敏感,ABA是参与植物抗逆生理过程的重要激素,对ABA敏感性较高的植物通常具有耐旱等特性。
2、转ZmbZIPa6南芥耐旱性鉴定
1)转ZmbZIPa6拟南芥苗期PEG模拟渗透胁迫的耐旱性鉴定:
将步骤一中T2代转ZmbZIPa6拟南芥株系1-6、2-4、3-3和野生型拟南芥Col-0(WT)的种子春化3d后播种于1/2MS培养基,培养2~3d后转移至含有用1/2MS+40%PEG浸泡48h的1/2MS培养基上,竖直培养,4周后观察表型,并拍照,且统计根长。
表型结果如图4中A所示,发现在培养4周后,野生型拟南芥Col-0(WT)全部死亡,而转ZmbZIPa6拟南芥株系1-6、2-4、3-3的存活率分别达到50%,83.33%和66.67%;根长的比较情况如图4中B所示,发现转ZmbZIPa6拟南芥株系1-6、2-4、3-3的根明显长于野生型拟南芥Col-0(WT),说明ZmbZIPa6能较好的抵抗PEG模拟的干旱胁迫。
2)转ZmbZIPa6拟南芥土壤干旱处理:
采用人工控水的方法进行土壤干旱,将步骤一中T2代转ZmbZIPa6拟南芥株系1-6、2-4、3-3和野生型拟南芥Col-0(WT)的种子4℃春化3d后,直接播在土中,幼苗长2周大小时,浇水至饱和,开始控水进行干旱处理。当处理至野生型拟南芥Col-0(WT)大部分出现严重失水萎蔫并叶片失绿时,立即恢复浇水,一周后调查统计干旱处理后的存活率(存活率=存活株数/试验总株数×100%)。
结果如图5所示,在连续控水干旱处理4周后,转ZmbZIPa6拟南芥株系1-6、2-4、3-3和野生型拟南芥Col-0(WT)生长均受到抑制,底部莲座叶开始出现萎蔫失绿现象。在浇水恢复一周后,部分转ZmbZIPa6拟南芥株系1-6、2-4、3-3植株能够存活并正常生长、结实;而全部野生型拟南芥Col-0(WT)植株萎蔫致死,复水后不能恢复生长,存活率为0%。
以上研究结果表明,转ZmbZIPa6拟南芥较野生型拟南芥Col-0具有较好的抗旱特性。
3、转ZmbZIPa6拟南芥耐碱性鉴定
将步骤一中T2代转ZmbZIPa6拟南芥株系1-6、2-4、3-3和野生型拟南芥Col-0(WT)的种子春化3d后播种于1/2MS培养基,培养2~3d后移至pH 9.0的1/2MS培养基上,并将平板竖直培养,2周后观察表型并拍照,且统计根长。
表型结果如图6中A所示,在培养2周后,转ZmbZIPa6拟南芥株系1-6、2-4、3-3和野生型拟南芥Col-0(WT)植株生长均受到抑制,而转ZmbZIPa6拟南芥株系1-6、2-4、3-3植株比野生型拟南芥Col-0(WT)植株的生长情况好;根长的比较情况如图6中B所示,转ZmbZIPa6拟南芥株系1-6、2-4、3-3植株与野生型拟南芥Col-0(WT)植株相比叶片大且根长,说明转ZmbZIPa6基因拟南芥具有良好的抗碱胁迫能力。
综上所述,ZmbZIPa6过表达转基因植株的耐旱性及耐碱性明显优于未转基因的植株,说明ZmbZIPa6是与植物耐旱及耐碱胁迫相关的蛋白。
以上对本发明进行了详述。对于本领域技术人员来说,在不脱离本发明的宗旨和范围,以及无需进行不必要的实验情况下,可在等同参数、浓度和条件下,在较宽范围内实施本发明。虽然本发明给出了特殊的实施例,应该理解为,可以对本发明作进一步的改进。总之,按本发明的原理,本申请欲包括任何变更、用途或对本发明的改进,包括脱离了本申请中已公开范围,而用本领域已知的常规技术进行的改变。按以下附带的权利要求的范围,可以进行一些基本特征的应用。
SEQUENCE LISTING
<110> 中国科学院植物研究所
<120> 转录因子ZmbZIPa6及其编码基因与应用
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<170> PatentIn version 3.5
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tgcagtctac tccagcctca ccgcctcagg tcggtcctac acggtcccgt tctgcctggt 60
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ctctctctcg tggcggcgcc tgggcagccg tcgctgccgc ctgcgctagg tgccaggtct 180
tcttcgggcc cttcgccggc gacgagtacc accaggttgt acatgtatac acccatgtgc 240
tttactgggt cggaccctac tcatcctcca cactaaggcc atctccatag actgcactgt 300
ttagactcca ctgtttagag agtggagata gggagtgaga tagagagtgg aatagggagg 360
atgctggaga tagcctaagg ttgccactga gtgtgtgcat gattcagagg caatgtcatc 420
acaaactgga ggtggcaaag agagtgacgc cggccctggg caacacaggc agatgcaaag 480
tttggcaaga caagggtctt tatacaacct taccctcgat gaggtgcaga accatttggg 540
ggagccattg cttagcatga attttgatga gctcctgaag agcgtgtttc ctgatggtgt 600
agattcagat ggtgctgtca caggcaagcc tgatcggacc tcgagcctgc agcgacaggg 660
gagcatcctg atgcctccgc agttgagcaa gaagactgtt gacgaggtgt ggaagggcat 720
ccagggtgga ccagagacca gtactgtggt ggatggtctg cagaggcgag agaggcatcc 780
aacacttggg gagatgacac ttgaggactt cttggtcaaa gctggggttg ttactgaagg 840
acttgtgaag gattcagctg attttccaag caatatggat acagctggca gcagtgttgt 900
tgtggctgct gcttctagtt tgaaccctgg agcacagtgg ttgcaacagt accagcagca 960
agttttgggg tcccagcaat tgagtttggc aggttcatac atggctagtc agttgcgccc 1020
tcagccattg tctattgcta caggtgctac tttggattca atttactctg atgaccagat 1080
tacttcgcca tcatttggtg cactttcgga ccctcagaca cctgggcgca agcgtggtgc 1140
cttaggggag gtagtggata aggtggttga aagaaggcag aagaggatga taaagaatag 1200
ggaatcagct gcgcgatcaa gagcgagaaa gcaggcttat actaatgaac ttgaaaacaa 1260
ggtattccgt ctagaagagg agaataagag gctaaagaaa cagcaggagt tggatgagat 1320
actgagctct gctcctcccc cagaacccaa atatcaactc aggagaacgg gttcggctgc 1380
cttttgagtt tcggcgctga caatgctcat agtgattttt gtacattata caaggattct 1440
actttatatg gtttccttcc ttttgtatta gaaacatttg atagattgag ggtatagctt 1500
tttcccggcc cggtaggtca gtgtatgtag gtcgctagca agctgtagca tgtcctggcg 1560
gatcatttcc ttggtgtctg tcagtgtgta catcttctag acacatgtct aagtctgaga 1620
gcttccttat acgagaaggt attcaggt 1648
<210> 2
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Gln His Arg Gln Met Gln Ser Leu Ala Arg Gln Gly Ser Leu Tyr Asn
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Leu Thr Leu Asp Glu Val Gln Asn His Leu Gly Glu Pro Leu Leu Ser
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Met Asn Phe Asp Glu Leu Leu Lys Ser Val Phe Pro Asp Gly Val Asp
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Ser Asp Gly Ala Val Thr Gly Lys Pro Asp Arg Thr Ser Ser Leu Gln
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Arg Gln Gly Ser Ile Leu Met Pro Pro Gln Leu Ser Lys Lys Thr Val
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Asp Glu Val Trp Lys Gly Ile Gln Gly Gly Pro Glu Thr Ser Thr Val
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Val Asp Gly Leu Gln Arg Arg Glu Arg His Pro Thr Leu Gly Glu Met
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Thr Leu Glu Asp Phe Leu Val Lys Ala Gly Val Val Thr Glu Gly Leu
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Val Lys Asp Ser Ala Asp Phe Pro Ser Asn Met Asp Thr Ala Gly Ser
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Ser Val Val Val Ala Ala Ala Ser Ser Leu Asn Pro Gly Ala Gln Trp
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Leu Gln Gln Tyr Gln Gln Gln Val Leu Gly Ser Gln Gln Leu Ser Leu
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Ala Gly Ser Tyr Met Ala Ser Gln Leu Arg Pro Gln Pro Leu Ser Ile
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Ala Thr Gly Ala Thr Leu Asp Ser Ile Tyr Ser Asp Asp Gln Ile Thr
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Ser Pro Ser Phe Gly Ala Leu Ser Asp Pro Gln Thr Pro Gly Arg Lys
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Lys Gln Ala Tyr Thr Asn Glu Leu Glu Asn Lys Val Phe Arg Leu Glu
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Glu Glu Asn Lys Arg Leu Lys Lys Gln Gln Glu Leu Asp Glu Ile Leu
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Ser Ser Ala Pro Pro Pro Glu Pro Lys Tyr Gln Leu Arg Arg Thr Gly
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Ser Ala Ala Phe
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<213> 人工序列(Artificial Sequence)
<400> 3
gtctcagaag accagagggc tattgagact tttcaacaaa gggtaatatc gggaaacctc 60
ctcggattcc attgcccagc tatctgtcac ttcatcgaaa ggacagtaga aaaggaagat 120
ggcttctaca aatgccatca ttgcgataaa ggaaaggcta tcgttcaaga atgcctctac 180
cgacagtggt cccaaagatg gacccccacc cacgaggaac atcgtggaaa aagaagacgt 240
tccaaccacg tcttcaaagc aagtggattg atgtgataac atggtggagc acgacactct 300
cgtctactcc aagaatatca aagatacagt ctcagaagac cagagggcta ttgagacttt 360
tcaacaaagg gtaatatcgg gaaacctcct cggattccat tgcccagcta tctgtcactt 420
catcgaaagg acagtagaaa aggaagatgg cttctacaaa tgccatcatt gcgataaagg 480
aaaggctatc gttcaagaat gcctctaccg acagtggtcc caaagatgga cccccaccca 540
cgaggaacat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat 600
gtgatatctc cactgacgta agggatgacg cacaatccca ctatccttcg caagaccctt 660
cctctatata aggaagttca tttcatttgg agaggacctc gagaaagagg atccacctga 720
ggcctccgtt ccatgggcta gaagcttctc ctcctctgct aacgtaagcc tctctgtttt 780
ttttctctgt ttcttttgaa atgaatccaa ttagtgatga taatctgtgt ttgatgtatc 840
attgatttaa catcttgaca atgaatcgtg atcggaagtg ataaagttat gggtcaacgg 900
tttcaaagag agagaaagac ttttagagtc aactctcgac tctttcttaa ttatgttatt 960
gctatttgtc tcttttcttg aagtctgaac aattcttggg attgttttgc aggttctagc 1020
ttctccaacc acaggaattc atcgcccggg actgggttat caacaagttt gtacaaaaaa 1080
gcaggctccg cggccgcccc cttcaccaga tcggctgagc caaggccatg tcatcacaaa 1140
ctggaggtgg caaagagagt gacgccggcc ctgggcaaca caggcagatg caaagtttgg 1200
caagacaagg gtctttatac aaccttaccc tcgatgaggt gcagaaccat ttgggggagc 1260
cattgcttag catgaatttt gatgagctcc tgaagagcgt gtttcctgat ggtgtagatt 1320
cagatggtgc tgtcacaggc aagcctgatc ggacctcgag cctgcagcga caggggagca 1380
tcctgatgcc tccgcagttg agcaagaaga ctgttgacga ggtgtggaag ggcatccagg 1440
gtggaccaga gaccagtact gtggtggatg gtctgcagag gcgagagagg catccaacac 1500
ttggggagat gacacttgag gacttcttgg tcaaagctgg ggttgttact gaaggacttg 1560
tgaaggattc agctgatttt ccaagcaata tggatacagc tggcagcagt gttgttgtgg 1620
ctgctgcttc tagtttgaac cctggagcac agtggttgca acagtaccag cagcaagttt 1680
tggggtccca gcaattgagt ttggcaggtt catacatggc tagtcagttg cgccctcagc 1740
cattgtctat tgctacaggt gctactttgg attcaattta ctctgatgac cagattactt 1800
cgccatcatt tggtgcactt tcggaccctc agacacctgg gcgcaagcgt ggtgccttag 1860
gggaggtagt ggataaggtg gttgaaagaa ggcagaagag gatgataaag aatagggaat 1920
cagctgcgcg atcaagagcg agaaagcagg cttatactaa tgaacttgaa aacaaggtat 1980
tccgtctaga agaggagaat aagaggctaa agaaacagca ggagttggat gagatactga 2040
gctctgctcc tcccccagaa cccaaatatc aactcaggag aacgggttcg gctgcctttt 2100
gagtctgtct agagtccgca aaaatcacca gtctctctct acaaatctat ctctctctat 2160
ttttctccag aataatgtgt gagtagttcc cagataaggg aattagggtt cttatagggt 2220
ttcgctcatg tgttgagcat ataagaaacc cttagtatgt atttgtattt gtaaaatact 2280
tctatcaata aaatttctaa ttcctaaaac caaaatccag tgacc 2325
Claims (10)
1.一种蛋白质,其特征在于,所述蛋白质为如下A1)或A2)或A3)任一所示:
A1)氨基酸序列如SEQ ID NO.2所示的蛋白质;
A2)SEQ ID NO.2所示的氨基酸序列的N端或/和C端连接蛋白标签得到的融合蛋白;
A3)将SEQ ID NO.2所示的氨基酸序列经过一个或几个氨基酸残基的取代和/或缺失和/或添加得到的与A1)所示的蛋白质具有90%以上的同一性且功能相同的蛋白质。
2.权利要求1所述蛋白质的编码基因。
3.根据权利要求2所述的编码基因,其特征在于:所述编码基因为如下B1)或B2)或B3)中任一所示:
B1)SEQ ID NO.1第413-1387位所示的DNA分子;
B2)编码序列为SEQ ID NO.1第413-1387位所示的DNA分子;
B3)在严格条件下与B1)或B2)限定的DNA分子杂交,且编码权利要求1所述蛋白质的DNA分子。
4.含有权利要求2或3所述的编码基因的表达盒、重组载体、重组微生物、转基因植物细胞系、转基因植物组织、转基因植物器官或转基因植株。
5.权利要求1所述的蛋白质或权利要求2或3所述的编码基因或权利要求4所述的表达盒、重组载体、重组微生物、转基因植物细胞系、转基因植物组织、转基因植物器官或转基因植株在下述任一中的应用:
C1)调控植物耐旱性;
C2)制备调控植物耐旱性的产品;
C3)提高植物耐旱性;
C4)制备提高植物耐旱性的产品;
C5)调控植物耐碱性;
C6)制备调控植物耐碱性的产品;
C7)提高植物耐碱性;
C8)制备提高植物耐碱性的产品;
C9)调控植物对ABA的敏感性;
C10)制备调控植物对ABA的敏感性的产品;
C11)提高植物对ABA的敏感性;
C12)制备提高植物对ABA的敏感性的产品。
6.权利要求1所述的蛋白质或权利要求2或3所述的编码基因或含有权利要求2或3所述的编码基因的表达盒、重组载体、重组微生物、转基因植物细胞系、转基因植物组织、转基因植物器官或转基因植株在植物育种中的应用。
7.一种培育耐旱性和/或耐碱性高的转基因植物的方法,其特征在于:所述方法包括提高目的植物中权利要求1所述的蛋白质的基因的表达量和/或所述蛋白质的含量和/或所述蛋白质的活性,得到转基因植物;所述转基因植物的耐旱性和/或耐碱性高于所述目的植物。
8.根据权利要求7所述的方法,其特征在于:所述提高目的植物中权利要求1所述蛋白质的基因的表达量和/或所述蛋白质的含量和/或所述蛋白质的活性的方法为在目的植物中表达或过表达权利要求1所述蛋白质。
9.根据权利要求8所述的方法,其特征在于:所述表达或过表达的方法为将权利要求1所述蛋白质的编码基因导入目的植物。
10.根据权利要求5或6所述的应用,或权利要求7-9任一所述的方法,其特征在于:所述植物是如下E1)至E4)中的任一种:
E1)双子叶植物;
E2)单子叶植物;
E3)十字花科植物;
E4)禾本科植物。
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2009073069A2 (en) * | 2007-10-31 | 2009-06-11 | Monsanto Technology, Llc | Genes and uses for plant enhancement |
CN104017061A (zh) * | 2013-03-01 | 2014-09-03 | 中国科学院植物研究所 | 转录因子ZmbZIP17及编码基因与其在响应逆境中的应用 |
US9290773B2 (en) * | 2002-10-02 | 2016-03-22 | Monsanto Technology Llc | Transgenic plants with enhanced agronomic traits |
-
2020
- 2020-06-04 CN CN202010499200.1A patent/CN113773374B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9290773B2 (en) * | 2002-10-02 | 2016-03-22 | Monsanto Technology Llc | Transgenic plants with enhanced agronomic traits |
WO2009073069A2 (en) * | 2007-10-31 | 2009-06-11 | Monsanto Technology, Llc | Genes and uses for plant enhancement |
CN104017061A (zh) * | 2013-03-01 | 2014-09-03 | 中国科学院植物研究所 | 转录因子ZmbZIP17及编码基因与其在响应逆境中的应用 |
Non-Patent Citations (3)
Title |
---|
KAIFA WEI等: "Genome-Wide Analysis of bZIP-Encoding Genes in Maize", DNA RESEARCH, vol. 19, pages 463 - 476, XP055460440, DOI: 10.1093/dnares/dss026 * |
LIRU CAO等: "Systematic Analysis of Dierentially Expressed Maize ZmbZIP Genes between Drought and Rewatering Transcriptome Reveals bZIP Family Members Involved in Abiotic Stress Responses", INT. J. MOL. SCI., vol. 20, pages 1 - 25 * |
NCBI: "ACCESSION NM_001156809:Zea mays uncharacterized LOC100283911 (LOC100283911), mRNA", Retrieved from the Internet <URL:https://www.ncbi.nlm.nih.gov/nuccore/226491957?sat=48&satkey=89617798> * |
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